Enhanced production of L-arginine by improving carbamoyl phosphate supply in metabolically engineered Corynebacterium crenatum

Wang, Qing; Jiang, Aiping; Tang, Jie; Gao, Hui; Zhang, Xian; Yang, Taowei; Xu, Zhenghong; Rao, Zhiming

doi:10.1007/s00253-021-11242-w

Cited by 11 publications

(5 citation statements)

References 46 publications

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“…In addition to the above result, the C. crenatum SYPA strain has another advantage as a putrescine producer that it carries a strong flux toward l -arginine formation . In natural organisms, putrescine is synthesized via the pathways of ADC and ODC.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to the above result, the C. crenatum SYPA strain has another advantage as a putrescine producer that it carries a strong flux toward L-arginine formation. 37 In natural organisms, putrescine is synthesized via the pathways of ADC and ODC. Therefore, the genes among the two pathways were heterologously expressed in C. crenatum SYPA for putrescine synthesis.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Production of Putrescine in Metabolic Engineering Corynebacterium crenatum by Mixed Sugar Fermentation

Yang

Qiu

Zhu

et al. 2022

ACS Sustainable Chem. Eng.

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Putrescine, a biogenic diamine, serves as an important component in various polyamides, medicine, and surfactants. One challenge for efficient putrescine production is to construct high-efficiency microbial cells. In this study, we designed the metabolic engineering strategies to significantly enhance the putrescine titer in Corynebacterium crenatum SYPA. First, an optimal synthetic pathway of putrescine comprising arginine decarboxylase (speA) and agmatinase (speB) genes from Escherichia coli was selected and introduced into C. crenatum. For efficient production of putrescine, the expression of agmatinase was optimized through a ribosome binding site regulation strategy. Next, the putrescine yield was furthermore increased by the knockout of snaA, speE, and cgmR to block putrescine degradation and overexpression of exporter CgmA. Additionally, the xylose utilization pathway was constructed for putrescine synthesis. Finally, the titer of putrescine was 41.5, 36.8, and 33.4 g/L from glucose, mixed sugar, and simulated wheat straw hydrolysates by fed-batch culture for 72 h, respectively. The yield was 0.18 g/g glucose, 0.15 g/g sugar, and 0.14 g/g sugar, respectively. To our knowledge, those results were the highest putrescine titers ever reported in the engineering of C. crenatum and Corynebacterium glutamicum. The engineering strains had the potential to produce putrescine from biomass hydrolysates.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

Production of Putrescine in Metabolic Engineering Corynebacterium crenatum by Mixed Sugar Fermentation

Yang

Qiu

Zhu

et al. 2022

ACS Sustainable Chem. Eng.

Self Cite

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“…Previous studies have demonstrated that overexpression of key enzymes can increase cellular amino acid production. 50 However, excessively strong expression can disrupt the tricarboxylic acid cycle and hinder product accumulation. 51 In this study, the gln A and aspC genes were initially overexpressed by replacing their native promoters with different strength promoters.…”

Section: Discussionmentioning

confidence: 99%

“…Gln, Asp, and CP play essential roles as precursors in the de novo synthesis of UTP. Previous studies have demonstrated that overexpression of key enzymes can increase cellular amino acid production . However, excessively strong expression can disrupt the tricarboxylic acid cycle and hinder product accumulation .…”

Section: Discussionmentioning

confidence: 99%

Multivariate Modular Metabolic Engineering for High Titer Uridine Triphosphate Production in Escherichia coli

Liu,

Chen,

et al. 2023

ACS Sustainable Chem. Eng.

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Uridine triphosphate (UTP) holds great potential for applications in the food and medicine fields. Compared with the chemical synthesis route, microbial biosynthesis of UTP through metabolic engineering approaches offers a sustainable and environmentally friendly alternative. However, the tightly regulated UTP metabolic pathway has presented challenges in building efficient microbial cell factories. To address this issue, we applied a modular metabolic engineering strategy to overcome four distinct bottlenecks in UTP production in Escherichia coli. First, by blocking the UTP catabolic module, we achieved a substantial accumulation of UTP up to 304.67 μM. Second, in the UTP synthetic module, a uridine 5′monophosphate (UMP) kinase variant (MtUMPK R141A/K148A ) was obtained based on crystal structure analysis, resulting in a 10.47-fold increase in catalytic capacity. Furthermore, by fusion expression of the UMP kinase variant and orotidine 5′phosphate decarboxylase, the conversion of UMP to UTP was improved, leading to UTP concentrations reaching 11.36 mM. Third, in the UTP precursor module, several key metabolism-related genes were engineered to enhance the supply of precursors, resulting in an 87.9% increase in UTP production. Finally, by implementing electron transport chain regulation with an ATP-sensing switch, the optimal strain E. coli K16 achieved an intracellular UTP concentration of 28.40 mM (13.75 g/L), which was 544.37-fold higher than that of the base strain. This study represents the design of a de novo UTP-producing strain through metabolic engineering, opening new avenues for UTP and its derivatives production.

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“…Since bacteria have strict regulation to ensure internal nitrogen homeostasis, the homeostasis of a nitrogen donor in an intracellular environment also provides a guarantee for efficient biosynthesis of value-added nitrogen-containing compounds. Even so, the intensity of endogenous or additional nitrogen supply is usually strengthened to further improve the production efficiency of microbial cell factories during synthesis process, for example, the supply of intracellular glutamine is increased by knocking out or overexpressing related genes to increase the titer of N -acetylglucosamine [93] or arginine [94] . In addition to metabolic engineering strategies that supplement endogenous nitrogen metabolism, external supplementation of nitrogen sources to adjust nitrogen content is also effective.…”

Section: Present and Potential Application For Synthetic Biologymentioning

confidence: 99%

Understanding and application of Bacillus nitrogen regulation: A synthetic biology perspective

He¹,

Li²,

Zhang³

et al. 2023

Journal of Advanced Research

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Enhanced production of L-arginine by improving carbamoyl phosphate supply in metabolically engineered Corynebacterium crenatum

Cited by 11 publications

References 46 publications

Production of Putrescine in Metabolic Engineering Corynebacterium crenatum by Mixed Sugar Fermentation

Production of Putrescine in Metabolic Engineering Corynebacterium crenatum by Mixed Sugar Fermentation

Multivariate Modular Metabolic Engineering for High Titer Uridine Triphosphate Production in Escherichia coli

Understanding and application of Bacillus nitrogen regulation: A synthetic biology perspective

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